title: Tetravalent antibody-scTRAIL fusion proteins with improved properties.
abstract: We applied the immunoglobulin E (IgE) heavy-chain domain 2 (EHD2) as the covalently linked homodimerization module to generate antibody-scTRAIL fusion proteins. By fusing a humanized single-chain fragment variable (scFv) directed against EGFR to the N-terminus of the EHD2 and a single-chain derivative of TRAIL (scTRAIL) to the C-terminus of the EHD2, we produced a dimeric, tetravalent fusion protein. The fusion protein retained its binding activity for EGFR and TRAIL receptors. In vitro, the targeted antibody-scTRAIL fusion protein exhibited an approximately 8- to 18-fold increased cytotoxic activity compared with the untargeted EHD2-scTRAIL fusion protein. This resulted in increased antitumor activity in a subcutaneous Colo205 xenograft tumor murine model. In summary, the scFv-EHD2-scTRAIL fusion protein combines target cell selectivity with an increased TRAIL activity leading to improved antitumor activities.
authors.affiliation: Corresponding Author: Roland E. Kontermann, Institut für Zellbiologie und Immunologie, Universität Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. roland.kontermann@izi.uni-stuttgart.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Seifert
authors.forename: Aline
authors.initials: A
authors.lastname: Plappert
authors.forename: Sina
authors.initials: S
authors.lastname: Fellermeier
authors.forename: Martin
authors.initials: M
authors.lastname: Siegemund
authors.forename: Klaus
authors.initials: K
authors.lastname: Pfizenmaier
authors.forename: Roland E
authors.initials: RE
authors.lastname: Kontermann
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology*; D017209:Apoptosis / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D051379:Mice; D009369:Neoplasms / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics* / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D053221:TNF-Related Apoptosis-Inducing Ligand; D007073:Immunoglobulin E; C512478:EGFR protein, human; D066246:ErbB Receptors